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Tomato Lectin Histochemistry for Microglial Visualization

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Microglia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1041))

Abstract

The use of different lectins for the study of microglial cells in the central nervous system (CNS) is a valuable tool that has been extensively used in the last years for the selective staining of this glial cell population, not only in normal physiological conditions, but also in a wide range of pathological situations where the normal homeostasis of the parenchyma is disturbed. In this chapter we accurately describe the methodology for the selective labelling of microglial cells by using the tomato lectin (TL), a protein lectin obtained from Lycopersicum esculentum with specific affinity for poly-N-acetyl lactosamine sugar residues which are found on the plasma membrane and in the cytoplasm of microglia. Here we describe how to perform this technique on vibratome, frozen, and paraffin sections for optical microscopy, as well as for transmission electron microscopy (TEM) studies. Using this methodology it is possible to visualize amoeboid microglia in the developing brain, ramified microglia in the adult, and activated/reactive microglia in the experimentally damaged brain. In addition, as TL also recognized sugar residues in endothelial cells, this technique is very useful for the study of the relationship established between microglia and the CNS vasculature.

In memoriam of Laia Acarin Pérez-Simó (1970–2011)

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Abbreviations

AAS:

Acetic acid solution

BBS:

Buffer blocking solution

CNS:

Central nervous system

DS:

Developing solution

DSN:

Developing solution with nickel

DSNC:

Developing solution with nickel-cobalt-enhanced DAB

EPBS:

Endogenous peroxidase blocking solution

FS:

Fixative solution

FSG:

Fixative solution with glutaraldehyde

GFAP:

Glial fibrillary acidic protein

GSA-IB4:

Griffonia simplicifolia B4 isolectin

MHC:

Major histocompatibility complex

ML-1:

Mistletoe lectin

OS:

Olmos solution

OTS:

Osmium tetroxide solution

PBS:

Phosphate buffer solution

PWB:

Phosphate washing buffer

RCA-1:

Ricinus communis agglutinin-1

SAS:

Sodium acetate solution

SS:

Sucrose solution

TB:

Tris-buffered solution

TBS:

Tris-buffered saline solution

TBST:

TBS with Triton X-100

TEM:

Transmission electron microscopy

TL:

Tomato lectin

UAS:

Uranyl acetate solution

WB:

Walpole buffer

WGA:

Wheat germ agglutinin

References

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Author information

Authors and Affiliations

  1. Universitat Autònoma de Barcelona, Barcelona, Spain

    Nàdia Villacampa, Beatriz Almolda & Berta González

  2. Universitat Autònoma de Barcelona, Bellaterra, Spain

    Bernardo Castellano

Authors
  1. Nàdia Villacampa
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  2. Beatriz Almolda
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  3. Berta González
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  4. Bernardo Castellano
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Editor information

Editors and Affiliations

  1. , Department of Oncology-Pathology, Karolinska Institutet, Solnavägen, Stockholm, SE-171 77, Sweden

    Bertrand Joseph

  2. , Department of Biochemistry, University of Seville, García González 2, Seville, 41012, Spain

    José Luis Venero

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© 2013 Springer Science+Business Media New York

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Villacampa, N., Almolda, B., González, B., Castellano, B. (2013). Tomato Lectin Histochemistry for Microglial Visualization. In: Joseph, B., Venero, J. (eds) Microglia. Methods in Molecular Biology, vol 1041. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-520-0_23

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  • DOI: https://doi.org/10.1007/978-1-62703-520-0_23

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-519-4

  • Online ISBN: 978-1-62703-520-0

  • eBook Packages: Springer Protocols

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